Week 3
From 2007.igem.org
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Alice.pasini (Talk | contribs) |
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::'''07/18/07''' | ::'''07/18/07''' | ||
| - | *We performed a test for GFP induction with IPTG. We picked a colony from the J04430 | + | *We performed a test for GFP induction with IPTG. We picked a colony from the J04430 and J04431 plates and grew it in 5 ml of LB medium during the day. In the afternoon we diluited the 5 ml cultures in 50 ml and let them growing overnight. |
*We picked a colony from 07/17 plates and inoculated in 5 ml of LB medium from each plate and incubated overnight. | *We picked a colony from 07/17 plates and inoculated in 5 ml of LB medium from each plate and incubated overnight. | ||
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::'''07/19/07''' | ::'''07/19/07''' | ||
| - | *We | + | *In the morning we diluited the J04430 and J04431 cultures to an OD=0.05. We grew the cultures till an OD = 0.4 – 0.6. We added 1 mM IPTG to induce GFP expression and we tested fluorescence after 2 h. (link results foto). |
| + | *Althought J04431 worked rigth in presence of IPTG, J04430 didn't. So, we performed a run on electroforesis gel for the J04430 plasmid. We found that it didn't match the rigth molecular weight. We thougth maybe there's something wrong with the plasmid. | ||
Revision as of 09:48, 23 August 2007
- 07/17/07
- We amplified some bio- bricks of interest for our project from the IGEM plates. We resuspended and transformed:
- J04430 to test the GFP fluorescence detection with our experimental set up;
- J04431 to test the GFP (+LVA) half-life since we needed a GFP with a short one for our project;
- J04500, the PLac promoter inducible by IPTG;
- J04631, the GFP (+LVA) protein.
- We straked on plates with the right antibiotic.
- 07/18/07
- We performed a test for GFP induction with IPTG. We picked a colony from the J04430 and J04431 plates and grew it in 5 ml of LB medium during the day. In the afternoon we diluited the 5 ml cultures in 50 ml and let them growing overnight.
- We picked a colony from 07/17 plates and inoculated in 5 ml of LB medium from each plate and incubated overnight.
- 07/19/07
- In the morning we diluited the J04430 and J04431 cultures to an OD=0.05. We grew the cultures till an OD = 0.4 – 0.6. We added 1 mM IPTG to induce GFP expression and we tested fluorescence after 2 h. (link results foto).
- Althought J04431 worked rigth in presence of IPTG, J04430 didn't. So, we performed a run on electroforesis gel for the J04430 plasmid. We found that it didn't match the rigth molecular weight. We thougth maybe there's something wrong with the plasmid.
- 07/20/07
- Plasmid digestion (link):we digested J04500 with Spe1 and Pst1 and J04631 with Xba1 and Pst1.
- We performed the gel extraction procedure and store at -20°C.
- We amplified amd transformed I13507 and R0051 from the IGEM plate.
